Hello netters!
I'm trying to overexpress two plant proteins in E.coli. I use the
expression vector pET3d and BL21(DE3) as E.coli strain. Both cDNAS
are cloned on the same strand having each a ribosome binding site (dicistron).
Thus, they are expected to be co-transcribed but translated separately
cf. the following scheme:
T7 Promoter ! RBS RBS
! / protein 1 / protein 2
5'===--------------===---------------- 3'
-------->
TRANSLATION
The problem:
Protein 2 is expressed but not protein 1. I use polyclonal antibodies
to monitor expression of each gene.
The question:
Can anyone help me to explain why protein 1 is not expressed?
I would appreciate all suggestions about.
Thanks in advance,
Georges,
E-mail: Jorge.hernandez at bota.unine.ch
°
° °
% ° ° °
%%% ° °
%%%%% Merry Christmas°°
%%%%%%%
T
I